摘要
尿道下裂是男性泌尿生殖系统常见的先天性畸形，发病率约为千分之一至千分之三，患者不能站立排尿，严重影响了患者的生活质量，手术重建尿道是最主要的治疗方法，目前手术方法众多，但是由于自体组织来源有限，效果并不理想，术后并发症较多，因此我们在动物实验的基础上于国内首次尝试应用同种异体脱细胞膀胱黏膜下基质来修复阴茎型尿道下裂以改善尿道下裂的临床治疗；同时鉴于大部分尿道下裂患者的病因不明，我们拟建立稳定可靠的尿道下裂动物模型以指导对尿道下裂病因学方面的研究。
人类尿道下裂的发病率近年来增长迅速，尿道下裂与环境污染的相关性不容忽视，目前随着工业化进程加速，全球化学合成物种类越来越多，环境污染日趋严重，各种生物变异现象层出不穷。许多化学合成物并未进行严格的致畸、致癌、致突变实验，生殖健康方面的影响更是无法预料。莠去津(除草剂)现在已被欧盟全面禁止，美国正在开展对它的应用评估，但是我国仍在大量供应和使用，其对人类的影响究竟如何，目前尚无定论，我们试图探讨莠去津是否具有类雌激素样作用、胚胎致畸作用及其对尿道下裂的发病率的影响，国内未见类似的报道。
一、脱细胞膀胱粘膜下基质尿道下裂修复的临床应用研究
我们采用反复冻融-酶法获得了脱细胞膀胱粘膜下基质，在口腔粘膜接种实验、大鼠腹部肌肉移植实验、新西兰白兔中段尿道缺损(1cm×0.5cm)修复等等实验的基础上于2004年将同种异体脱细胞膀胱黏膜下基质试用于3例阴茎型尿道下裂患者的治疗。
结果：
1)HE染色显示脱细胞膀胱粘膜下基质采用反复冻融-酶法脱细胞完全，Masson染色染色组织学观察显示脱细胞基质的主要成分为胶原，采用冷冻干燥技术处理脱细胞膀胱粘膜下基质得到冻干材料，组织学和扫描电镜观察证实其具有三维立体网络状结构。
2)2例一期手术患者尿线粗、直，无尿瘘，无散射：术后1年随访，阴茎无下弯，无尿痛、尿瘘，尿线粗，尿流测定仪及尿道镜检查未见异常。另1例二期手术患者，电话随访远端尿道未见狭窄及闭锁。
结论如下：
1．反复冻融-酶法和冷冻干燥技术处理膀胱粘膜下层可以获得冻干脱细胞膀胱粘膜下基质，呈三维立体网络状结构，组织相容性好，能够在新生组织形成过程中逐渐降解吸收，符合组织工程支架材料的要求。
2．应用同种异体脱细胞膀胱粘膜下基质重建阴茎远端型尿道下裂的尿道，术后1年近期效果满意，表明同种异体脱细胞膀胱粘膜下基质是一种有前途的较为理想的尿道组织修复替代材料，远期效果还有待继续进行严密仔
Hypospadias is one of the most common congenital deformity in males with a reported incidence of 3/1000 live male births. There is an increasing tendence in the incidence rate of hypospadias in many countries. The cause of most cases of hypospadias is unclear. The relating studies showed that the pathogenies of hypospadias were related to genetic factors and environmental factors.
Recent epidemiologic studies suggest that parents working in areas of high pesticide application are at increased risk for adverse reproductive outcomes such as infertility ,poor fertilization, fetal death and congenital abnomalies .Residential pesticide exposures and their effects on reproductive health or sexual mal-differentiation are less-well understood.
In the recent years, a great deal of attentive has focused on the global presence of endocrine-disrupting contaminants in the environment .
Atrazine(2-chloro-4-ethylamino-6-isopropylamine-1,3,5-triazine) is the most commonly used herbicide in the US and our country. The US Department of Agriculture reports that more than 30,000 tons are used annually in the US alone ,and currently Atrazine is used in more than 80 countries .It is applied to control weeds in corn and sorghum fields ,both prior to and following emergence ,As a result of this widespread use , Atrazine is the most frequently detected pesticide in ground and surface water ,Therefore ,humans and wildlife are at risk for exposure to Atrazine.
Atrazine has also been reported to affect endocrine function in the male. Some scholars reported it can reduce production of testosterone in juvenile rats .Atrazine, a triazine herbicide marketed by Ciba-Geigy throughout the United Stated and our country, but was forbidded in Europen Union. It is unknown whether Atrazine can induce hypospadias in animal experiment. So we
引文
1．曹谊林，崔磊，刘伟．中国组织工程研究回顾及发展．中华实验外科杂志，2003，20：485-6．
2. Langer RS, Vacanti JP. Tissue engineering. Science, 1994, 260: 920-6.
3. Meezan E, Hjelle JT, Brendel K. A simple, versatile, nondisruptive method for the isolation of morphologically and chemically pure basement membranes from several tissue. Life Sci, 1975, 17: 1721-32.
4. Probst M, Dahiya R, Carrier S, et al. Reproduction of functional smooth muscle tissue and partial bladder replacement. Br J Urol, 1997, 79: 505-15.
5. Wilson GJ, Courtman DW, Klement P, et al. Acellular matrix: a biomaterial approach for coronary artery bypass and heart valve replacemant. Ann Thorac Surg, 1995, 60: S353-9.
6. Courtman DW, Pereica CA, Kashef V, et al. Development of a pericardical acellular matrix biomaterial:Biochemical and mechanical effect of cell extraction. J Biomed Mater Res, 1994, 28: 655-66.
7. Courtman DW, Errett BF, Wilson GJ. The role of cross linking in modification of the immune response elicited against xenogenic vascular acellular matrices. J Biomed Mater Res, 2001, 55: 576-86.
8. Livesey SA, Herndon DN, Hollyoak MA, et al. Transplanted acellular allograft dermal matrix. Potential as a template for the reconstruction of viable dermis. Transplant, 1995, 60: 1-9.
9. Wainwright DJ. Use of an acellular allograft dermal matrix (Alloderm) in the management of full-thickness burns. Burns. 1995; 21: 243-8.
10. Takami Y, Matsuda T, Yoshitake M, et al. Dispase/detergent treated dermal matrix as a dermal substitute. Burns, 1996, 22: 182-90.
11. Walter RJ, Matsuda T, Reyes HIM, et al.. Characterization of acellular dermal matrices (ADMs) prepared by two different methods. Burns, 1998, 24: 104-13.
12 邹忠桃，谭谦．脱细胞异种真皮皮下埋植的实验研究．东南大学学报(医学版)，2003，22：148-50．
13. Dahms SE, Piechota H J, Dahiya R, et al. Composition and biomechanical properties of the bladder acellular matrix graft: comparative analysis in rat, pig and human. Br Jurol, 1998, 82: 411-9.
14. Voytik-Harbin SL, Brightman AO, Krajne MR, et al. Identification of extractable growth factors from small intestinal submucosa. J Cell Biochem, 1997; 67: 478-91.
15．王彦君，孔维佳，毕胜斌等．自制天然软骨细胞外基质支架材料的生物相容性研究．临床耳鼻咽喉科杂志，2004，18：363-366．
16．李海宁，王春仁，曹红英等．T-1型脱细胞异体组织补片生物相容性评价研究．中国医疗器械杂志，2004，28：117-119．
17．章庆国，赵士芳，林鹤等．超低温冻存同种异体成骨细胞与异种脱钙骨复合异位组织工程化骨．第二军医大学学报，2003，24：784-787．
18．孙新君，王正国，朱佩芳等．不同处理异种骨植入后受体T淋巴细胞亚型变化．中国临床康复，2004，8：2044-2045．
19. Allman A J, McPherson TB, Badylak SF, et al. Xenogeneic extracellular matrix grafts elicit a TH_2-restricted immune response. Transplantation, 2001, 71: 1631-40.
20. Raeder RH, Badylak SF, Sheehan C, et al. Natural anti-galactose alphal, 3 galactose antibodies delay, but do not prevent the acceptance of extracellular matrix xenografts. Transpl Immunol, 2002, 10: 15-24.
21．顾建军，陈家祺，彭鸿钧等．脱细胞真皮与异体巩膜在眼睑重建中的实验研究．眼科研究，2003，21：229-233．
22．左金华，杨佑成，李武修等．脱细胞异体真皮基质生物学特性的实验研究．临床口腔医学杂志，2004，20：276-278．
23．龙莉，吴春根，潘銮凤等．心脏瓣膜组织工程支架的制备及细胞种植．生物医学工程学杂志，2004，21：610-613．
24 刘太华，张炎，姜宗来等．组织工程血管脱细胞支架主要组织相容性复合体Ⅰ抗原性的免疫组化观察．第二军医大学学报，2004，25：72-74．
25. DeSagun EZ, Botts JL, Srivastava A, et al. Long-term outcome of xenogenic dermal matrix implantation in immunocompetent rats. J Surg Res,. 2001, 96: 96-106.
26. Badylak SF, Kropp B, McPherson T, et al. Small intestinal submucosa: a rapidly resorbed bioscaffold for augmentation cystoplasty in dog model. Tissue Eng, 1998, 4: 379-87.
27. Record RD, Hillegonds D, Simmoms C, et al. In vivo degradation of ~(14)C-labeled small intestinal submucosa (SIS) when used for urinary bladder repair. Biomaterials, 2001, 22: 2653-9.
28. Brendel K, Duhamel RC, Malone JM, et al. Acellalar vascular matrix as a small diamater vascular graft. In Kambic H, Kantrowitz A, Surg P. Vascular graft update: Safety and performance, Philadephia, ASTM, 1986.
29. Wilson GJ, Courtman DW, Klement P, et al. Acellular matrix allograft small caliber vascular prosthesis. Trans Am Soc Artif Intern Organs, 1990, 36: 340-3.
30. Wilson GJ, Courtman DW, Klement P, et al. Acellular matrix: a biomaterial approach for coronary artery bypass and heart valve replacemant. Ann Thorac Surg, 1995, 60: S353-9.
31. Courtman DW, Errett BF, Wilson GJ. The role of cross linking in modification of the immune response elicited against xenogenic vascular acellular matrices. J Biomed Mater Res, 2001, 55: 576-86.
32. Bader A, Schilling T, Teebken OE, et al. Tissue engineering of heart valves: Human endothelial cell seeding of detergent acellularized porcine valves. Eur J Cardiothorac Surg, 1998, 14: 279-84.
33. Zelinger J, Landeen LK, Alexander HG, et al. Development and characterization of tissue-engineered aortic valves. Tissue Engineering, 2001, 7: 9-22.
34．陶凯，毛天球，杨维东等．骨髓基质成骨细胞-松质骨基质复合人工骨修复颅骨缺损的实验研究，中华整形外科杂志，2002，18：226-8．
35．张晨，金铸，高景恒等．同种异体兔耳软骨ECM复制兔耳软骨的实验研究．实用美容整形外科杂志，1997，8：228-31．
36．张晨，马晓光，于鹏等．软骨脱细胞基质的研制及其力学性质分析．实用美容整形外科杂志，2002，13：59-62．
37．栾杰，蔡哲，杨佩瑛等．软骨细胞在异体脱细胞软骨基质上的体外培养实验．中华整形烧伤外科杂志，1999，15：178-9．
38. Fansa H, Keihoff G, Wolf K, et al. Tissue engineering of peripheral nerves: a comparison of venous and acellular muscle grafts with cultured schwann cells. Plast Reconstr Surg, 2001, 107: 485-97.
39．张思胜，刘世清，彭吴等．脱细胞异体神经修复鼠坐骨神经缺损．广东医学，2004，25：22-24
40. Kropp BP, Rippy MK, Badylak SF, et al. Regenerative urinary bladder augmentation using small intestinal submucosa: urodynamic and histopathologic assessment in long-term canine bladder augmentations. J Urol, 1996, 155: 2098-104.
41. Vaught JD, Kropp BP, Sawyer BD, et al. Detrusor regeneration in the rat using porcine small intestinal submucosal grafts: functional innervation and receptor expression. J Urol, 1996, 155: 374-8.
42. Clarke KM, Lantz GC, Salisbury SK, et al. Intestine submucosa and polypropylene mesh for abdominal wall repair in dogs.J Surg Res,1996, 60: 107-14.
43. Prevel CD, Eppley BL, Summerlin DJ, et al. Small intestinal submucosa: utilization as a wound dressing in full-thickness rodent wound. Ann Plast Surg, 1995, 35: 381-8.
44. Cobb MA, Badylak SF, Janas W, et al. Porcine small intestinal submucosa as a dural substitute. Surg Neurol, 1999, 51: 99-104.
45．顾延，戴克戎．猪小肠粘膜下层替代兔跟腱的实验研究．中华医学杂志，2002，82：1279-1282
46. Badylak SF, Lantz GC, Coffey A, et al. Small intestine submucous as a large diameter vascular graft in the dog.J Sur Res, 1989, 47: 74-80.
47. Lantz GC, Badylak SF, Coffey AC, et al. Small intestinal sabmucosa as a small diameter arterial graft in the dog. J Invest Surg, 1990, 3: 217-27.
48. Lantz GC, Badylak SF, Coffey AC. Small intestinal submucosa as a superior vena cava graft in the dog. J Surg Res, 1992, 53: 175-81.
49. Sandusky GE, Lantz GC, Badylak SF. Healing comparison of small intestine submucosa and ePTFE grafts in the canine carotid artery. J Surg Res, 1995, 58: 415-20.
50. Matheny RG, Hutchison ML, Dryden PE, et al. Porcine small intestinesubmacosa as a pulmonary valve leaflet substitute. J Heart Valve Dis, 2000, 9: 769-74.
51. Yoo JJ, Meng J, Oberpenning F, et al. Bladder augmentation using allogenic submucosa seeded with cells. Urology, 1998, 51: 221-5.
52. Sutherland RS, Baskin LS, Hayward SW, et al. Regeneration of bladder urothelium, smooth musle, blood vessels and nerves into an acellular tissue matrix. J urol, 1996, 156: 571-7.
53. Dahms SE, Piechota HJ, Dahiya R, et al. Composition and biomechanical properties of the bladder acellular matrix graft: comparative analysis in rat, pig and human. Br Jurol,1998, 82: 411-9.
54．刘流，梁德江，申鹏飞等．脱细胞膀胱细胞外基质作为膀胱替代材料的实验研究．中华实验外科杂志，1999，16：181-2．
55. Badylak SF. Xenogeneic extracellular matrix as a scaffold for tissue reconstruction. Transpl Immunol. 2004, 12: 367-77.
56. Livesey SA. US Patent. #5, 336, 616 1994; lifecell Corporation, 23-4.
57. Wainwright DJ. Use of an acellular allograft dermal matrix (Alloderm) in the management of full-thickness burns. Burns. 1995; 21:243-8.
58. Takami Y, Matsuda T, Yoshitake M, et al. Dispase/detergent treated dermal matrix as a dermal substitute. Burns, 1996, 22:182-90.
59. Rennekampff HO, Kiessig V, Govindaraj S, et al. Acellular human dermis promotes cultured keratinocyte engraftment. J Burn Care Rehabil, 1997, 18: 535-44.
60．霍孟华，戚可名，黄金井等．脱细胞异体真皮基质皮下移植后胶原的动态变化．中华整形外科杂志，2004，20：51-52
61. Achauer BM, VanderKamVM, Celikoz B, et al. Augmentation of facial soft tissue defects with Alloderm dermal graft. Ann Plast Surg, 1998, 41:503-7.
62. Kridel RWH, Foda H, Lunde KC. Septal perforation repair with acellular dermal allograft. Arch Otolargngol Head Neck Surg, 1998, 124:73-8.
63. Romo Ⅲ T, Sclafani AP, Falk AN, et al. A graduated approach to the repair of nasal septal perforations. Plast Reconstr Surg, 1999,103:66-75.
64. Izumi K,Takacs G, Terashi H, et al. Ex vivo development of a composite human oral mucosal equivalent. J Oral Maxillofac Surg, 1999, 57: 571-7.
65．法永红，李志韧，蔡兴伟等．异体软组织修复材料在口腔颌面部缺损中的应用．现代口腔医学杂志，2004，18：67-68
66. Tobin HA, Karas ND. Lip augmentation using an Alloderm graft. J OralMaxillofac Surg, 1998, 56: 722-7.
67. Novae AB Jr, Grisi DC, Molina GO, et al. Comparative 6-month clinical study of subepithelial connective tissue graft and acellular dermal matrix graft for the treatment of gingival recession. J Periodontol, 2001, 72: 1477-84.
68. McFeely WJ Jr, Bojrab DI, Kartush JM. Tympanic membrane perforation repair using Alloderm. Otolaryngol Head Neck Surg, 2000, 123: 17-21.
69. Inoue Y, Anthony JP, Lleon P, et al. Acellular human dermal matrix as a small vessel substitute. J Reconstr Microsurg, 1996, 12: 307-11.
70. Isch JA, Engum SA, Ruble CA, et al. Patch esophagoplasty using Alloderm as a tissue scaffold. J Pediatr Surg, 2001, 36: 266-8.
71．柴岗，曹谊林，尹健．骨组织工程应用研究进展．实用美容整形外科杂志，2002，13：264-5．
72．张旗涛，姚猛，于友等．同种异体脱细胞骨的生物力学性质及其临床应用．中华小儿外科杂志，2004，25：172-175．
73. Tajima K, Tohyama K, Ide C, et al. Regeneration through nerve allografts in the cynomolgus monkey. J Bone Joint Surg Am, 1991, 73: 172-85.
74. Dumont CE, Hentz VR. Enhancement of axon growth by detergent-extracted nerve grafts. Transplantation, 1997, 63: 1210-5.
75. Kim JC, Tseng SC. Transplantation of preserved human amniotic membrane for surface reconstruction in severely damaged rabbit corneas. Cornea, 1995, 14: 473-84.
76. Avila M, Espana M, Moreno C. et al. Reconstruction of ocular surface with heterologous limbal epithelium and amniotic membrane in a rabbit model. Cornea, 2001, 20: 414-20.
77. Matsumoto T, Holmes RH, Burdick CO, et al. Replacement of large veins with free inverted segments of small bowel: Autografts of submucosal membranes in dogs and clinical use. Ann Surg, 1966,164: 845-8.
78. Kropp BP, Ludlow JK, Spicer D, et al. Rabbit urethral regeneration using small intestinal submucosa onlay grafts. Urology. 1998, 52: 138-42.
79. Chen F, Yoo JJ, Atala A.: Acellular collagen matrix as a possible "off the shelf"biomaterial for urethral pepair. Urology, 1999, 53: 407.
80. Chen F, Yoo J J, Atala A. Experimental and clinical experience using tissue regeneration for urethral reconstuction. World J Urol, 2000, 18: 67-70.
81. Sievert KD, Bakircioglu ME, Nunes L, et al. Homologous acellular matrix graft for urethral reconstruction in the rabbit: histological and functional evaluation. J Urol, 2000, 163: 1958-65.
82. Parnigotto PP, Gamba PG, Conconi MT, Midrio P.: Experimental defeat in rabbit urethra repaired with acellular aortic matrix. Urol Res, 2000, 28: 46-61.
83. Sievert KD, Wefer J, Bakircioglu, et al. Heterologous acellular matrix graft for reconstruction of the rabbit urethra: histological and functional evaluation. J urol, 2001, 165: 2096-102.
84. Filippo RE, Yoo J J, Atala A.: Urethral replacement using cell seeded tubularized collagen matrices. J Urol, 2002, 168: 1789
85．刘流，梁德江，申鹏飞等．异体真皮细胞外基质重建尿道的实验和临床研究．中华泌尿外科杂志，2001，22：428-31．
86. Probst M, Dahiya R, Carrier S, et al. Reproduction of functional smooth muscle tissue and partial bladder replacement. Br J Urol, 1997, 79: 505-15.
87. Atala A, Guzman LF, Retik AB.: A novel inert collagen matrix for hypospadias repair. J Urol, 1999, 162: 1148-51.
88. EL-Kassaby AW, Retik AB,Yoo JJ, et al. Urethral stricture repair with off-the-shelf collagen matrix. J Urol, 2003, 169: 170-3.
1. Piechota HJ, Dahms SE, Nanes LS, et al. In vitro functional properties of the rat bladder regenerated by the bladder acellular matrix graft. J Urol, 1998, 159: 1717-1724.
2．邹忠桃，谭谦．脱细胞异种真皮皮下埋植的实验研究．东南大学学报(医学版)，2003，22：148-150．
3．杨光辉，崔磊，刘伟，等．组织工程技术修复全层皮肤缺损的研究．中华实验外科杂志，2003，20，408-410．
4. Meezan E, Hjelle JT, Brendel K. A simple, versatile, nondisruptive method for the isolation of morphologically and chemically pure basement membranes from several tissue. Life Sci, 1975, 17: 1721-1732.
5．温玉麟主编．药物与化学物质毒性数据．天津：天津科学技术出版社，1989：386．
6．天然细胞外基质制备技术．见杨志明主编．组织工程．北京：化学工业出版社，2002：215-227．
7．傅洪滨，霍孟华，张云涛．鼠异体无细胞真皮基质(ADM)的制备和应用．中国烧伤创疡杂志，2000；4：18～21．
8. Vacanti CA, Kim W, Upton J, et al. Tissue-engineered growth of bone and cartilage. Trans Proc, 1993, 25: 1019-1021.
9. Vacantino JP. Selective cell transplantation using bioabsorbable artificial polymers as matrices. J Pediatric Surg, 1988, 23:3-9.
10. Kropp BP, Ludlow JK, Spicer D, et al. Rabbit urethral regeneration usingsmall intestinal submucosa onlay grafts. Urology. 1998, 52: 138-142.
11. Cilento BG, Retik AB, Atala A. Urethral reconstruction using a polymer mesh. J Urol, suppl, 1995, 153: 371.
12. Bergman S, Javadpour N, Wade C, Terrill R.: Biodegradable ureteral grafts in dogs. Investigative Urol, 1978, 16:48.
13. Tachibana M, Nagamatsu GN, Addonizio JC.: Ureteral replacement using collagen sponge tube grafts. J Urol, 1985, 133: 866-869.
14. Bach AD, Bannasch H, Galla TJ, Bittner KM, Stark GB.: Fibrin glue as matrix for cultured autologous urothelial cells in urethral reconstruction. Tissue Engineering, 2001, 7: 45-53.
15. Romagnoli G, De Luca M, Faranda F.: Treatment of posterior hypospadias by the autologous graft of cultured urethral epithelium. New Engl J Med, 1990, 323: 527-530.
16. Romagnoli G, De Luca M, Faranda F.: One-step treatment of proximal hypospadias by the autologous graft of cultured urethral epithelium. J Urol, 1993, 150: 1204-1207.
17．刘流，梁德江，申鹏飞等．异体真皮细胞外基质重建尿道的实验和临床研究．中华泌尿外科杂志，2001，22：428-431．
18. Probst M, Dahiya R, Carrier S, et al. Reproduction of functional smooth muscle tissue and partial bladder replacement. Br J Urol, 1997, 79: 505-515.
19. Atala A, Guzman LF, Retik AB.: A novel inert collagen matrix for hypospadias repair. J Urol, 1999, 162: 1148-1151.
20. EL-Kassaby AW, Retik AB,Yoo JJ, et al. Urethral stricture repair with off-the-shelf collagen matrix. J Urol, 2003, 169: 170-173.
21．刘立强．中国协和医科大学博士研究生学位论文．2004．
22．王永前．中国协和医科大学博士研究生学位论文．2004．
23．王永前，李养群，刘立强等．冻干无细胞膀胱黏膜下基质修复兔尿道缺损．中华整形外科杂志，2005，21：62-65．
24. Duckett JW.: MAGPI(meatal advancement and glanuloplasty): a procedure for subcoronal hypospadias. Urol Clin N Amer, 1981, 8: 513-519.
25．张涤生，黄文义，徐春阳．前尿道延伸术一期修复下裂．中华外科杂志，1983，21(6)：362-3．
26．刘勇刚．前尿道延伸术一期矫治阴茎型尿道下裂(附22例报告)．广西医学，2000，22：976-977．
27. Mathieu P. Traitement en un temps de 1 hypospadiasbalanique' etjuxta-balanique.J Chi(Paris), 1932,39:481.
28. Edmond T, Gonzales JR, Krishna A.: The management of distal hypospadias with meatal-based vascularized flaps. J Urol, 1983,129:119-120.
29. Mastarde JC.One-stage correction of distal hypospadias and other people's fistulae.Br J Plast Surg, 1965,18:413.
30. Devine C Jr, Horton C.A one-stage hypospadias repair. J Urol, 1961,85:166.
31. Horon CE, Devine CJ.: Hypospadias In Converse(ed).: Reconstr Plast Surg, 1977,7:3845-3881.
32. Duckett JW. The island flap technique for hypospadias repair. Urol Clin North Am, 1981,8:503-511.
33. Klijn AJ, Dik P, de Jong TP. Results of preputial reconstruction in 77 boys with distal hypospadias. J Urol 2001 Apr;165(4):1255-1257.
34. Imamoglu MA, Bakirtas H, Tuygun C, et al. Clinical experiences with different one-staged surgical methods for primary hypospadias cases. Int Urol Nephrol. 2002; 33(1):107-112.
35. Elder JS, Duckett JW, et al. Onlay island flap in the repair of mid and distal penile hypospadias without chordee. J Urol, 1987,138:376-379.
36. DucketJW. Current hypospadias techniques. Can J Urol, 1995, 2(Suppl):30-32.
37. Snodgrass W, Patterson K, Plaire JC, et, al. Histology of the urethral plate: implications for hypospadias repair. J Urol, 2000,164(3 Pt 2):988-989.
1. Baskin LS. Hypospadias and urethral development. J Urol 2000, 163(3): 951-956.
2．吴艳乔，王艳萍，朱军，等．国人尿道下裂的流行病学特征．中华泌尿外科杂志，1997，18(12)：753-757．
3. PaulozziLJ, Erickson JD, Jackson RJ. Hypospadias trends in two US surveillance systems. Pediatrics, 1997, 100(5): 831-834.
4. Gray LE Jr, Ostby J, Monosson E, et al. Environmental antiandrogens: low doses of the fungicide vinclozolin alter sexual differentiation of the male rat. Toxicol Ind Health, 1999, 15(1-2): 48-64.
5. Mylchreest E, Wallace DG, Cattley RC, et, al. Dose-dependent alterations in androgen-regulated male reproductive development in rats exposed to Di(n-butyl) phthalate during late gestation. Toxicol Sci, 2000, 55(1): 143-151.
6. Gray LE Jr, Ostby J, Furr J, et al. Perinatal exposure to the phthalates DEHP, BBP, and DINP, but not DEP, DMP, or DOTP, alters sexual differentiation of the male rat. Toxicol Sci, 2000, 58(2): 350-365.
7. Miyagawa S, Buchanan DL, Sato T, et al. Characterization of diethylstilbestrol-induced hypospadias in female mice. Anat Rec, 2002, 266(1): 43-50.
8. Kojima Y, Hayashi Y, Mizuno K, et al. Spermatogenesis, fertility and sexual behavior inahypospadiac mouse model. J Urol, 2002, 167(3): 1532-1537.
9. Bowman CJ, Barlow NJ, Turner KJ, et al. Effects ofin utero exposure tofinasteride on androgen-dependent reproductive development in the male rat. Toxicol Sci, 2003, 74(2): 393-406.
10. Mylchreest E, Sar M, Cattley RC, et, al. Disruption of androgen-regulated male reproductive development by di(n-butyl)phthalate during late gestation in rats is different from flutamide. Toxicol Appl Pharmacol, 1999, 156(2): 81-95.
11. Kurzrock EA, Jegatheesan P, Cunha GR, et al. Urethral development in the fetal rabbit and induction of hypospadias: a model for human development. J Urol, 2000, 164(5): 1786-1792.
12. McIntyre BS, Barlow N J, Foster PM. Androgen-mediated development in male rat offspring exposed to flutamide in utero: permanence and correlation of early postnatal changes in anogenital distance and nipple retention with malformations in androgen-dependent tissues. Toxicol Sci, 2001, 62(2): 236-249.
13. Higuchi TT, Palmer JS, Gray LE Jr, et al. Effects of dibutyl phthalate in male rabbits following in utero, adolescent, or postpubertal exposure. Toxicol Sci, 2003, 72(2): 301-313.
14．贺厚光，张炜，朱佳庚．小鼠尿道下裂动物模型的建立．中华男科学2004，10(3)：172-174．
15．龚春雨，黄鲁刚，李虹等．已烯雌酚建立尿道下裂大鼠模型的实验研究．中华小儿外科杂志，2004，25：180-181
16. Baskin LS. Hypospadias and urethral development. J Vrol, 2000, 163: 954-955
17．陈荣月，母义明．非那雄胺的药理学及其临床应用．国外医学·药学分册，1994，21：356-360
18．刘孝东，李虹，步宏等．非那雄胺对大鼠前列腺微血管的影响．四川大学学报·医学版 2004，35：226-228
19. Shono T, Ramm Anderson S, Goh DW, et al. The effect of flutamide on testicular descent in rats examined by scanning electronmicroscopy. Pediatr Surg, 1994, 29(6): 839 844.
20. Koff SA, Jayanthi VR. Preoperative treatment with human chorionic gonadotropin in infancy decreases the severity of proximal hypospadias and chordee. J Urol, 1999, 162(4): 1435-9.
21. Forest MG, David M, de Peretti E, et al. Disorders of sexual differentiation in infants andchildren. Ann Endocrinol, 1980, 41(6): 545-53.
22. Knorr D, Beckmann D, Bidlingmaier F, Plasma testosterone in male puberty. Ⅱ. hCG stimulation test in boys with hypospadias. Acta Endocrinol, 1979, 90(2): 365-71.
23. Kiely EA, Chapman RS, Bajoria SK, et al. Maternal serum human chorionic gonadotrophin during early pregnancy resulting in boys with hypospadias or cryptorchidism. Br J Urol, 1995, 76(3): 389-92.
24. Feyaerts A, Forest MG, Morel Y, et al. Endocrine screening in 32 consecutive patients with hypospadias. J Urol, 2002, 168(2): 720-5.
25. Walsh PC, Curry N, Mills RC, et al. Plasma androgen responce to hCG stimulation in prepubertal boys with hypospadias and cryptorchidism. J Clin Endocrinol Metab, 1976, 42(1): 52-9.
26. Arnold S. F. Synergistic activation of estrogen receptor with combinations of environmental chemicals. Science, 1996, 272: 1489-1492.
27. Bsuer SB, Retik AB, Colodny SH. Genetic aspects of hypospadias. Urol Clin N Am, 1981, 8: 559—564.
28．杨东仁，何凤生，付桂平．我国农药使用品种现状分析．中华劳动卫生职业病杂志，1998，16：348-350．
29．张宏军．加拿大削减莠去净的使用．农药科学管理，2004，25(3)：12
30．刘亚萍．美国环保局维持莠去净使用．登记农药科学毒理，2004，25(4)：41-42．
31．范国珍．莠去净的药害问题及药害防范技术研究概述．农药科学与管理，2003，24(1)：20-23．
32. Wetzel LT, Luempert Ⅲ LG, Breckenridge CB, et al. Chronic effects of atrazine on estrus and mammary tumor formation in female Sprague-Dawley and Fischer 344 rats. J Toxicol Environ Health, 1994; 43(2): 169-182.
33. Eldridge JC, Wetzel LT, Stevens JT, et al. The mammary tumor response in triazine-treated female rats: a threshold-mediated interaction with strain and species-specific reproductive senescence. Steroids, 1999; 64(9): 672-678.
34. Cooper RL, Stoker TE, Tyrey L, et al. Atrazine disrupts the hypothalamic control of pituitary-ovarian function. Toxicol Sci, 2000; 53(2): 297-307.
35. Larson DL, McDonald S, Fivizzani AJ, et al. Effects of the herbicide atrazine on Ambystoma tigrinum metamorphosis: duration,larval growth, and hormonal response, Physiol Zool, 1998, 71: 671-679.
36. Grady J, Wieser C, Wiebe J, et al. An evaluation of atrazine as a potential endocrine disruptor in Largemouth Bass. In: Proceedings of SETAC 19th Annual Meeting. PMPO13, 1996.
37. Andrew S Friedmann. Atrazine inhibition of testosterone prodution in rat males following peripubertal exposure. Reproductive Toxicology, 2002, 16: 275-279.
38. R Infurna, B Levy, C Meng et al. Teratological evaluations of atrazine technical, a triazine herbicide, in rats and rabbits. Journal of Toxicology and Environmental Health, 1988, 24: 307-319.
39．陈海燕，王心如．环境雌激素的识别与评价．中国工业医学杂志，2001，14：369—373．
40．刘兆平．环境雌激素活性物质的检测．国外医学卫生学分册，2000，27(2)：96—100．
41. Korach KS, McLachlan JA. Techniques for detection of estrogenicity. J Environ Health Perpect, 1995, 103(S7): 5-8.
42. Nishikawa J, Saito K, Goto J, et al. New screening methods for chemicals with hormonal activities using interaction of nuclear hormone receptor with coactivator. Toxicol Appl Pharmacol, 1999, 154(1): 76-83.
43. Anderson HR, wersson AM, Arndd SF, et al. Comparison of short term estrogenicity tests for identification of hormone-disrupting chemicals. J Environ Health Perspect, 1999, 107(supple 1): 89—108
44. John Doull. Proceedings of 5th International congress of Pesticide chemistry. Pergamen Press, 1982: 433-436.
45．杜克久，徐晓白．环境雌激素研究进展．科学通报，2000，45：2241—2251